Abstract: The present invention includes the characterization of the DGK? gene and the generation of screening methods for compounds that inhibit the function of DGK?. The DGK family of enzymes occupies a signaling crossroads since they catalyze the phosphorylation of DAG to produce PA. Both the substrate (DAG) and the product (PA) of this reaction are key factors in intracellular signaling, making the regulation of DGK? activity important to understand and control. DGK? ?/? mice were also generated and studied to assist in understanding the function of DGKs in regulating cellular signaling. DGK? displays selectively for 20:4-DAG and is highly expressed in different areas of the brain, including Purkinje cells in the cerebellum, hippocampal interneurons, and the Pyramidal neurons in the CA3 region of the hippocampus.

Abstract: The present invention provides a method of screening for agents which may regulate or inhibit the activities of TACE and TGF-?. These agents may act by enhancing or inhibiting the activity of DGK-?. An activity of DGK-? is the enzymatic conversion of DAG to PA. The method includes contacting a cell or organism with a test agent determining the activity of DGK-? within the cell. An agent with reduces or increases the activity of DGK may be used to regulate or inhibit the activity of TACE and TGF-?. Such agents can be used to treat diseases such as cancer where cell growth and division and inflamation are important factors.

Abstract: The present invention includes the characterization of the DGKe gene and the generation of screening methods for compounds that inhibit the function of DGKe. The DGK family of enzymes occupies a signaling crossroads since they catalyze the phosphorylation of DAG to produce PA. Both the substrate (DAG) and the product (PA) of this reaction are key factors in intracellular signaling, making the regulation of DGKe activity important to understand and control. DGKe −/− mice were also generated and studied to assist in understanding the function of DGKs in regulating cellular signaling. DGKe displays selectively for 20:4-DAG and is highly expressed in different areas of the brain, including Purkinje cells in the cerebellum, hippocampal interneurons, and the Pyramidal neurons in the CA3 region of the hippocampus.

Abstract: The present invention provides a method of screening for agents which may regulate or inhibit the activities of TACE and TGF-&agr;. These agents may act by enhancing or inhibiting the activity of DGK-&dgr;. An activity of DGK-&dgr; is the enzymatic conversion of DAG to PA. The method includes contacting a cell or organism with a test agent determining the activity of DGK-&dgr; within the cell. An agent with reduces or increases the activity of DGK may be used to regulate or inhibit the activity of TACE and TGF-&agr;. Such agents can be used to treat diseases such as cancer where cell growth and division and inflamation are important factors.

Abstract: Two novel diacylglycerol kinase (DGK) isoforms are disclosed. The cDNA of one DGK isoform, designated DGK&egr;, is about 2.6 kb in length. The DGK&egr; cDNA has an open reading frame encoding 567 amino acids and has a predicted molecular mass of 63 kDA. DAG kinase &egr; is highly selective for arachidonate-containing diacylglycerol (DAG) substrates. The cDNA of the second isoform, designated DGK &zgr;, is 3.5 kb in length. The DGK&zgr; cDNA contains a single large open reading frame encoding a 928-amino acid protein with a predicted molecular mass of 103.9 kDa. An alternatively spliced muscle specific species of DGK&zgr;, DGK&zgr;-2 is also disclosed and characterized. DGK&zgr; is localized to the nucleus. A lysine-rich region with homology to the MARCKS protein is shown to be necessary and sufficient to confer nuclear localization to DGK&zgr; via a protein kinase C phosphorylation.

Abstract: Two novel diacylglycerol kinase (DGK) isoforms are disclosed. The cDNA of one DGK isoform, designated DGK.epsilon., is about 2.6 kb in length. The DGK.epsilon. cDNA has an open reading frame encoding 567 amino acids and has a predicted molecular mass of 63 kDA. DAG kinase .epsilon. is highly selective for arachidonate-containing diacylglycerol (DAG) substrates. The cDNA of the second isoform, designated DGK .zeta., is 3.5 kb in length. The DGK.zeta. cDNA contains a single large open reading frame encoding a 928-amino acid protein with a predicted molecular mass of 103.9 kDa. An alternatively spliced muscle specific species of DGK .zeta., DGK.zeta.-2 is also disclosed and characterized. DGK.zeta. is localized to the nucleus. A lysine-rich region with homology to the MARCKS protein is shown to be necessary and sufficient to confer nuclear localization to DGK.zeta. via a protein kinase C phosphorylation.